• Title/Summary/Keyword: DNA fingerprinting

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GENOTYPING OF STREPTOCOCCUS MUTANS USING AP-PCR IN CHILDREN WITH RAMPANT CARIES (AP-PCR을 이용한 다발성 우식아동의 구강내 Streptococcus mutans의 유전자형 분류)

  • Jang, Myung-Jo;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.65-81
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    • 1997
  • For the purpose of evaluating the appropriateness of AP-PCR as a facile, rapid and reproducible method for genotyping Streptococcus mutans, and selecting the discriminant primer for it, a DNA fingerprinting was performed on the microorganisms isolated from caries-free children and children with rampant caries, respectively. In the course of selecting appropriate primer for S. mutans genotyping, we chose S2 primer from 6 different primers which shows highest resolution on the agarose gel as well. Nineteen kinds of fingerprint patterns were observed in caries-free children and children with rampant caries which were produced by combination of 7 different fragments. Interestingly, the number of types observed in caries-free children was greater than that in children with rampant caries. And we observed Type 2 was predominant in children with rampant caries (about 80%) and relatively even distribution of each types in caries-free children. Furthermore, it was appeared that the major types in normal control were not or rarely found in children with rampant caries. In conclusion, we could establish simple, rapid and highly reproducible AP-PCR method for genotyping S. mutans. We also found differences in distribution of S. mutans between normal and patient, which suggested that cariogenicity is also dependent on qualitative aspects which is caused by the difference in genotypes of S. mutans in oral cavity.

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Recurrent Pseudomonas aeruginosa Infection in Chronic Lung Diseases: Relapse or Reinfection?

  • Yum, Ho-Kee;Park, I-Nae;Shin, Bo-Mun;Choi, Soo-Jeon
    • Tuberculosis and Respiratory Diseases
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    • v.77 no.4
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    • pp.172-177
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    • 2014
  • Background: Pseudomonas aeruginosa infection is particularly associated with progressive and ultimately chronic recurrent respiratory infections in chronic obstructive pulmonary disease, bronchiectasis, chronic destroyed lung disease, and cystic fibrosis. Its treatment is also very complex because of drug resistance and recurrence. Methods: Forty eight cultures from 18 patients with recurrent P. aeruginosa pneumonia from 1998 to 2002 were included in this study. Two or more pairs of sputum cultures were performed during 2 or more different periods of recurrences. The comparison of strains was made according to the phenotypic patterns of antibiotic resistance and chromosomal fingerprinting by pulsed field gel electrophoresis (PFGE) using the genomic DNA of P. aeruginosa from the sputum culture. Results: Phenotypic patterns of antibiotic resistance of P. aeruginosa were not correlated with their prior antibiotic exposition. Fifteen of 18 patients (83.3%) had recurrent P. aeruginosa pneumonia caused by the strains with same PFGE pattern. Conclusion: These data suggest that the most of the recurrent P. aeruginosa infections in chronic lung disease occurred due to the relapse of prior infections. Further investigations should be performed for assessing the molecular mechanisms of the persistent colonization and for determining how to eradicate clonal persistence of P. aeruginosa.

Susceptibility for breast cancer in young patients with short rare minisatellite alleles of BORIS

  • Yoon, Se-Lyun;Kim, Dae-Cheol;Cho, Se-Heon;Lee, Sang-Yeop;Chu, In-Sun;Heo, Jeong-Hoon;Leem, Sun-Hee
    • BMB Reports
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    • v.43 no.10
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    • pp.698-703
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    • 2010
  • In this study, we characterized two blocks of minisatellites in the 5' upstream region of the BORIS gene (BORIS-MS1, -MS2). BORIS-MS2 was found to be polymorphic; therefore, this locus could be useful as a marker for DNA fingerprinting. We assessed the association between BORIS-MS2 and breast cancer by a case-control study with 428 controls and 793 breast cancers cases. Rare alleles in the younger group (age, <40) were associated with a statistically significant increased risk of breast cancer (odds ratio, 4.84; 95% confidence interval, 1.06-22.22; and P = 0.026). A statistically significant association between the short rare alleles and cancer was identified in the younger group (8.02; 1.01-63.83; P = 0.021). Kaplan-Meier estimates showed that poor prognosis was associated with patients who contained the rare alleles. Our data suggest that the short rare alleles of BORIS-MS2 could be used to identify the risk for breast cancer in young patients.

Molecular fingerprinting of olive flounder pathogenic Streptococcus parauberis strains by random amplified polymorphic DNA analysis

  • Jung, Yong-Uk;Kang, Sang-Hyuck;Jin, Chang-Nam;Kang, Bong-Jo;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.899-903
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    • 2005
  • Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

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Genetic Diversity of the Original Plant for Taraxaci Herba, Taraxacum spp. by the Analysis of AFLP (AFLP 분석을 통한 포공영 기원식물 민들레의 유전 다양성 분석)

  • Kim, Wook Jin;Moon, Byeong Cheol;Ji, Yunui;Lee, Young Mi;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.4
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    • pp.247-254
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    • 2013
  • Collected germplasms of five representative dandelion species (Taraxacum ohwianum, T. platycarpum, T. platypecidum, T. officinale, and T. coreanum) were 104 lines from different habitates in Korea and China. Their genetic diversity was analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 1,176 total DNA fragments and 523 polymorphic bands with a 44.4% ratio of polymorphism. On the basis of similarity coefficient analysis by unweight pair group method with arithmetic averages (UPGMA), 104 dandelion germplasm lines were ranged from 0.64 to 0.99 and clustered distinct five group depending on the species. Furthermore, a principal coordinate analysis (PCA) by the application of multi-variate analysis indicated significantly greater differences among species than geographical origins.

Molecular typing of epidemiologically unrelated Staphylococcus epidermidis recovered from dogs by pulsed-field gel electrophoresis

  • Pak, Son-il
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.811-818
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    • 1999
  • A total of 16 Staphylococcus epidermidis isolates collected from 14 dogs admitted to the Veterinary Medicial Teaching Hospital in Seoul National University over eleven months were examined for in vitro antibiotic susceptibility pattern with minimum inhibitory concentration (MIC) and slime production, a virulence-associated phenotype, and were genetically characterized by pulsed-field gel electrophoresis (PFGE). The frequency of resistance to antimicrobial agents tested was not high, with a susceptibility ranging from 56.3% to 100%. Three strains exhibited multiple drug resistance against amikacin (MIC, $32-64{\mu}g/ml$), ampicillin ($32{\mu}g/ml$), fosfomycin ($32-128{\mu}g/ml$) and gentamicin ($16{\mu}g/ml$). Vancomycin, ciprofloxacin and rifampin were effective antibiotics against the isolates. All isolates were slime producers ; strains isolated from dogs which died of bacteremia were more likely to produce slime than those isolated from dogs which survived. Chromosomal DNA fingerprinting of the isolates yielded 16 different genomic types with few common bands, indicating a variety of clones of S epidermidis were prevalent in the hospital. This study revealed that PFGE is an useful method for the genotype characterization of S epidermidis strains and this organism could probably be pathogenic in some dogs with severe disorders. Further works on a larger number of epidemiologically defined strains are required to assess these results.

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Quantification and Graphical Method for DNA Fingerprinting (유전자검사자료의 통계분석을 위한 수량화 및 그래프 방법)

  • 박미라
    • The Korean Journal of Applied Statistics
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    • v.15 no.1
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    • pp.85-105
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    • 2002
  • To explore the relationships among frequencies for sets of alleles, within or between loci, is one of the first analyses in population genetic study. The general question is whether the frequency of a set of alleles is the same as the product of each of the separate allele frequencies. For two alleles of a single locus, Hardy-Weinberg equilibrium is tested and for an allele from each of two loci, linkage disequilibrium is tested. However, it is more useful if we can quantify and graphically represent this information. In this study, we suggest graphical methods to find associations between alleles. We also analyze the STR data of Korean population as an illustration.

Molecular Typing of Vibrio parahaemolyticus by Repetitive Element-PCR (rep-PCR) (Repetitive Element-PCR (rep-PCR)을 이용한 Vibrio parahaemolyticus 의 분자유전학적 아형 분류)

  • Kim, Won Sik;Hong, Seung Bok;Lee, Kyung;Lee, Jung Nam;Shin, Kyeong Seob
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.1
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    • pp.1-6
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    • 2004
  • The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

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Genetic Diversity of Curcuma Genus Collected Germplasm using Analysis of AFLP (AFLP 분석을 통한 Curcuma속 수집종간 유전적 다양성 분석)

  • Moon, Byeong Cheol;Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.6
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    • pp.455-460
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    • 2013
  • Collected germplasms of five representative species belonging to Curcuma genus (C. longa, C. aromatica, C. zedoaria, C. phaeocaulis and C. kwangsiensis) were 52 samples from different farmhouse in Korea and China. To elucidate the genetic diversity among the species, 52 samples were analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 643 total DNA fragments and 349 polymorphic bands with the 54.3% ratio of polymorphism. In the analysis of coefficient similarity using unweight pair group method with arithmetic averages (UPGMA), 52 Curcuma germplasm lines were ranged from 0.60 to 0.99 and clustered distinct five groups according to the species and collected geographical levels. However, the result of principal coordinate analysis (PCA) by multi-variate analysis was shown significantly greater differences among species than geographical origins based on AFLP profiling data of these samples.

PCR-mediated Fingerprinting to Identify Dang-Gui(당귀) (당귀류 한약재의 유전자 감별 연구)

  • 최호영;정유헌;고지완
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.11-15
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    • 2000
  • Radix Angelicae Gigantis is sweet and pungent in flavor, warm in property. Its effects are tonifying the blood, promoting blood circulation, relieving pain and moistening the bowels. Its indications are blood deficiency syndrome characterized by sallow complexion, dizziness, irregular menstruation, amenorrhea, pains due to blood stasis, and rheumatic arthralgia. Using genes of A. gigas, A. acutiloba, and A. sinensis, the origin of which is identified, as criteria, we analysed many kinds of Angelica with RAPD and RFLP on ITS region, in order to compare and discriminate genes extracted from crude drugs ‘Dang-gui’, that are produced in Korea on the one hand and imported on the other hand. We reached the following conclusion. 1. We could extract DNA from both original plant and dried plant. 2. Especially Uniprimer #1, Uniprimer #2, Uniprimer #4 and Uniprimer #9 were useful. 3. Among the restriction enzymes Sma I, Msp I, Hae III, and Hinf I, used in this experiment, four restriction enzymes except Hinf I could be used properly in discriminating all samples used as A. gigas. We think that this result can be used as a method of discriminating crude drug of Angelica L. related drugs, and used in controlling quality and circulation.

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